GSM7179003: IC-ATGR1, rep1; Homo sapiens; RNA-Seq
Total RNA was extracted from 4-6 organoids derived from ACE-/- (clone AC1), AGTR1-/- (clone AT1) and Isogenic Control (IC-ACE and IC-AGTR1) iPSC lines using the Quick-RNA MiniPrep kit (Zymo Research, R1054) per the manufacturer instructions. RNA samples from organoids derived from n=3 independent differentiation experiments from each line were quantified using Qubit® RNA BR Assay kit. Normalized in nuclease free water to 8 ng/uL and run on Agilent Fragment Analyzer for peak analysis. RQN values averaged 8.6 with a range of 7.4-9.9. 28S/18S ratio averaged 1.85 with a range of 1.6 to 2.2. 100 ng of RNA were prepared into libraries following Illumina Stranded Total RNA Prep with Ribo-Zero Plus protocol with all recommended reagents. Final library amplification step used 2.5 uL index anchors and 14 PCR cycles. Individual libraries were quantified using Qubit® dsDNA HS assay and run on the Agilent Fragment Analyzer for library bp size. Libraries were pooled using the percent range adjusted concentration of fragments 175-650 bp. Library pool was run on 1.5% Bio-Rad Certified Low Range Ultra Agarose for size selection greater than 150 bp. DNA library was purified following MinElute Gel Extraction Kit Protocol.